ICCVAM Recommendations for Use of the LLNA for Evaluating the... of Pesticide Formulations and Other Products

ICCVAM Recommendations for Use of the LLNA for Evaluating the Allergic Contact Dermatitis Potential
of Pesticide Formulations and Other Products
1U.S.
ICCVAM has updated its 1999 validation report on the LLNA based on a recent evaluation of the
usefulness and limitations of the LLNA for assessing the skin sensitizing potential of pesticide
formulations. This review was initiated because the original report did not include an analysis of
the LLNA for these types of substances, and there were growing regulatory concerns that the
LLNA might not identify sensitizing pesticide formulations. LLNA data from 104 formulations were
included in the evaluation, most of which are water soluble and therefore were tested in an
aqueous vehicle containing 1% Pluronic L92. Of the pesticide formulations for which LLNA and
guinea pig data were available (n=23), the LLNA classified 52% (12/23) as sensitizers, while GP
tests classified only 13% (3/23) as sensitizers. All three of the pesticide formulations identified as
sensitizers in the GP test were also identified as sensitizers in the LLNA; there were no instances
of underprediction by the LLNA. Thus, there appears a greater likelihood of obtaining a positive
result in the LLNA than in a GP test. These studies also provide data for aqueous solutions that
emphasize the need for careful selection of an appropriate vehicle that maintains test substance
contact with the skin (e.g., 1% Pluronic L92 in water) to achieve adequate exposure when testing
such substances. Based on these data, ICCVAM agreed with an international peer review panel
that the LLNA could be used for testing pesticide formulations, and products in aqueous vehicles,
unless there are physicochemical properties that may interfere with the ability of the LLNA to
detect the sensitizing potential of a substance. ICCVAM recommendations will be forwarded to
Federal agencies for regulatory acceptance consideration. Adoption of these recommendations
should expand the use of the LLNA for skin sensitization testing, thereby reducing and refining
animal use for this purpose.
J Matheson1, A Jacobs2, M Wind1, J Chen3, M Hashim3, M Lewis3, E Margosches3, D McCall3, T McMahon3, J Redden3, R Ward3, W Stokes4
Consumer Product Safety Commission, Bethesda, MD; 2U.S. Food and Drug Administration, Silver Spring, MD; 3U.S. Environmental Protection Agency, Washington, DC;
4National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods, Research Triangle Park, NC
• 
Table 1: Summary of LLNA Performance for Testing Pesticide
Formulations and Other Products, Metal Compounds, and
Substances in Aqueous Solutions
Comparison
LLNA vs.
Reference Test
Method Results
n1
LLNA vs. GP3
25
Accuracy
%
Sensitivity
No.2
%
No.2
LLNA False
Negative
Rate
%
No.2
Specificity
• ICCVAM concludes that these data support the usefulness of the LLNA for testing
pesticide formulations and other products, metals (with the exception of nickel), and
substances tested in aqueous solutions, unless there are unique physiochemical
properties associated with these materials that may interfere with the ability of the LLNA
to detect sensitizing substances.
LLNA False
Positive Rate
%
No.2
%
No.2
52
11/21
48
10/21
− 
Substances Tested in Aqueous Solutions
56
14/25
75
3/4
25
1/4
− 
Pesticide Formulations
LLNA vs. GP3
23
57
13/23
100
3/3
0
0/3
50
10/20
50
10/20
0/9
60
3/5
40
2/5
−
Metal Compounds
LLNA vs.
GP3
14
86
12/14
LLNA vs. Human4
6
83
5/6
LLNA vs. Human4
12
42
5/12
100
100
9/9
5/5
0
0
0/5
0
0/1
100
1/1
25
2/8
75
6/8
−
Natural Complex Substances
75
3/4
25
1/4
• 
When testing aqueous formulations in the LLNA, an appropriate vehicle should be
added to prevent the test substance from running off the skin (e.g. added pluronic
acid to achieve 1% Pluronic L92 [Boverhof et al. 2008]) so an adequate dermal
exposure is achieved.
If an LLNA variant (e.g., a nonradioactive LLNA version) is validated for use to test
novel substance classes, then the findings should be relevant to the family of
validated and accepted LLNA tests.
• 
• The Interagency Coordinating Committee on the
Validation of Alternative Methods (ICCVAM) is
charged with evaluating the scientific validity of new,
revised, and alternative toxicological test methods
applicable to U.S. Federal agency safety testing
requirements (Sailstad et al. 2001).
− ICCVAM forwards recommendations to Federal
agencies.
− By law, the agencies must respond to ICCVAM
within 180 days.
• In response to a nomination by the U.S. Consumer
Product Safety Commission in 2007, ICCVAM
evaluated the applicability domain of the murine local
lymph node assay (LLNA), a test method for
assessing the potential of substances to cause
allergic contact dermatitis (ACD).
− ACD is an allergic skin reaction characterized by redness, swelling, and itching that can
result from repeated contact with a sensitizing chemical or product.
•  ICCVAM’s recommendations regarding the use of the LLNA for testing pesticide formulations
and other products, metals, and substances in aqueous solutions (i.e., the current
applicability domain of the LLNA) are documented in a Test Method Evaluation Report
(TMER).
−
The ICCVAM TMER includes recommendations regarding:
  Current usefulness and limitations of the LLNA
  An LLNA test method protocol
  Future studies
• The information summarized in this poster is based on
a retrospective review of LLNA data derived from a
database of over 600 substances (including pesticide
formulations and other products) and builds on the
1998 ICCVAM evaluation of the LLNA (ICCVAM 1999;
Dean et al. 2001; Haneke et al. 2001), which
considered LLNA data for 211 substances.
• Table 1 shows LLNA accuracy statistics compared to
guinea pig and human results for the products and
substances considered in this evaluation, which were
derived from the database described above.
• The remainder of this poster will focus on the
evaluation of pesticide formulations and substances
tested in aqueous solutions.
Environmental Protection Agency
Office of Pesticide Programs
Jonathan Chen, Ph.D.
Masih Hasim, D.V.M., Ph.D.
Marianne Lewis
Deborah McCall
Timothy McMahon, Ph.D.
John Redden
Jenny Tao, Ph.D.
Office of Prevention, Pesticides,
and Toxic Substances
Elizabeth Margosches, Ph.D.
Ronald Ward, Ph.D.
Office of Research and Development
Marsha Ward, Ph.D.
Office of Science Coordination and Policy
Karen Hamernik, Ph.D.
Food and Drug Administration
Center for Devices and Radiological
Health
Vasant G. Malshet, Ph.D., DABT
Jeffrey Toy, Ph.D.
Center for Drug Evaluation and Research
Paul Brown, Ph.D.
Abigail Jacobs, Ph.D. (IWG Co-chair)
Jiaqin Yao, Ph.D.
Center for Veterinary Medicine
Ruth Barratt, Ph.D., D.V.M.
National Institute of Environmental
Health Sciences
Dori Germolec, Ph.D.
William Stokes, D.V.M., DACLAM
National Institute for Occupational Safety
and Health
B. Jean Meade, D.V.M., Ph.D.
European Centre for the Validation of
Alternative Methods - Liaison
Silvia Casati, Ph.D.
Alexandre Angers, Ph.D.
Japanese Center for the Validation of
Alternative Methods - Liaison
Hajime Kojima, Ph.D.
33
2/6
40
2/5
60
3/5
0
0/1
100
Figure 1: Schematic of LLNA test method protocol
Apply Test Substance
Administer Radioisotope
(3H or 125I)
•
•
•
•
The LLNA database included 171 studies representing 139 substances.
− The substances were tested in the LLNA at a final concentration of at least 20% water.
− 91 substances (123 LLNA studies) were pesticide formulations and pure compounds.
− 75 substances were pesticides tested in aqueous 1% Pluronic L92.
− 48 substances (48 LLNA studies) were aqueous eluates of medical devices.
GP data were available for 25 substances tested in aqueous solutions.
− The LLNA and the GP results were in agreement (accuracy) 56% (14/25) of the time
(Table 1).
−  11 substances were discordant between the LLNA and the GP tests.
−  10/11 discordant substances were pesticide formulations tested in aqueous 1%
Pluronic L92; these were the same 10 substances discussed for the pesticide
formulations analysis, and all were overpredicted by the LLNA with respect to the GP
results (48% [10/21] false positive rate) (Table 1).
34% (25/75) pesticide formulations tested in aqueous 1% Pluronic L92 produced negative
results in the LLNA.
Neomycin sulfate, tested in 25% ethanol, was underpredicted by the LLNA with respect to
the GP (25% [1/4] false negative rate) (Table 1).
Because of sample preparation differences between the pesticide formulations and pure
compounds, and the medical device eluates, these groups were analyzed separately.
− All 48 medical device eluates were LLNA negative (no GP data were available).
−  These eluates were not analyzed to determine their constituents, or whether any
compound(s) were eluted from the medical devices.
Days 1 - 3
Day 6
5 hours
Measure Proliferation
(Scintillation Counts)
SI =
Prepare Single
Cell Suspension
Mean DPM of Treatment Group
Mean DPM of Control Group
Collect Draining
Auricular Lymph
Nodes
SI ! 3 = Sensitizer (Positive)
SI < 3 = Nonsensitizer (Negative)
2. Pesticide Formulations
•
•
•
The updated LLNA database included data for 104 pesticide formulations.
23 formulations had LLNA and GP data for the same formulation.
There were no human skin sensitization test data or post-marketing sensitization report
data.
For the 23 formulations with both GP and LLNA data:
−  LLNA and the GP results were in agreement (accuracy) 57% (13/23) of the time
(Table 1).
− All 3 pesticide formulations identified as sensitizers in the GP test were also identified
as sensitizers in the LLNA.
− The LLNA classified 52% (12/23) of formulations as sensitizers while GP tests
classified 13% (3/23) as sensitizers.
− The LLNA identified 7 additional substances as sensitizers that were classified as
nonsensitizers in GP tests, an overprediction (i.e., false positive) rate of 50% (10/20)
(Table 1).
− No pesticide formulations were underpredicted (i.e., false negative) by the LLNA
compared to the guinea pig results.
!
Event
January 10, 2007
CPSC nominates six LLNA review activities for ICCVAM evaluation,1 including the
LLNA applicability domain.
January 2007
ICCVAM IWG is re-established to work with NICEATM to carry out LLNA
evaluations.
January 24, 2007
ICCVAM endorses the CPSC-nominated LLNA review activities.
May 17, 2007
Federal Register notice (72 FR 27815) – The Murine Local Lymph Node Assay:
Request for Comments, Nominations of Scientific Experts, and Submission of Data
June 12, 2007
SACATM endorses with high priority the six CPSC-nominated LLNA review activities.
January 8, 2008
Federal Register notice (73 FR 1360) – Announcement of an Independent Scientific
Peer Review Panel Meeting on the Murine Local Lymph Node Assay; Availability of
Draft Background Review Documents; Request for Comments
March 4–6, 2008
International Independent Scientific Peer Review Panel convenes in public session
with opportunity for oral public comments at CPSC Headquarters in Bethesda, MD,
to review new versions and applications of the LLNA.
May 20, 2008
Federal Register notice (73 FR 29136) – Announcement of the Peer Review Panel
Report on the Validation Status of New Versions and Applications of the Murine
Local Lymph Node Assay (LLNA): A Test Method for Assessing the Allergic Contact
Dermatitis Potential of Chemicals and Products: Notice of Availability and Request
for Public Comments2
June 18–19, 2008
SACATM public meeting: comments on the 2008 Panel report
February 27,
2009
Federal Register notice (74 FR 8974) – Announcement of a Second Meeting of the
Independent Scientific Peer Review Panel on the Murine Local Lymph Node Assay;
Availability of Draft Background Review Documents (BRD); Request for Comments
April 28–29, 2009
International Independent Scientific Peer Review Panel convenes in public session
with opportunity for oral public comments, at NIH Natcher Conference Center in
Bethesda, MD, to review new versions and applications of the LLNA.
1. Substances Tested in Aqueous Solutions
•
June 1, 2009
Federal Register notice (74 FR 26242) – Independent Scientific Peer Review Panel
Report: Updated Validation Status of New Versions and Applications of the Murine
Local Lymph Node Assay: A Test Method for Assessing the Allergic Contact
Dermatitis Potential of Chemicals and Products: Notice of Availability and Request
for Public Comments3
June 25–26, 2009
SACATM public meeting: comments on the 2009 Panel report
October 28, 2009
ICCVAM endorses TMER for the LLNA applicability domain, which includes LLNA
Addendum on the validity of the LLNA for mixtures, metals, and aqueous solutions.
Abbreviations: BRD = Background Review Document; CPSC = U.S. Consumer Product Safety Commission; ICCVAM
= Interagency Coordinating Committee on the Validation of Alternative Methods; IWG = ICCVAM Immunotoxicity
Working Group; LLNA = Murine Local Lymph Node Assay; NICEATM = National Toxicology Program Interagency
Center for the Evaluation of Alternative Toxicological Methods; NIH = National Institutes of Health; SACATM =
Scientific Advisory Committee on Alternative Toxicological Methods; TMER = Test Method Evaluation Report.
ICCVAM-recommended future studies include:
− To more comprehensively evaluate the ability of the LLNA to be used for testing nickel
compounds, additional data from LLNA studies on such compounds with comparative
human and/or GP data are needed.
−  Available solubility data should be provided so that thermodynamic activity can be
computed and compared to maximum theoretical percutaneous penetration.
  Consider this information when comparing LLNA data from studies in lipophilic
delivery systems vs. aqueous systems.
− Use 1% Pluronic L92 in water as the vehicle for aqueous formulations in order to
expand the existing database for that vehicle, unless adequate scientific rationale is
provided for using another aqueous vehicle.
−  For new classes of test materials, conduct an integrated assessment of available
information, including:
  Computer-assisted structure-activity relationships
  Prediction/measurement of biotransformation to potential reactive species
  Possibly peptide, protein, or lipid binding
− While recommending future studies, ICCVAM emphasizes avoidance of revalidation of
the LLNA for new classes/types of test substances unless a biologically-based rationale
exists.
− Before conducting animal testing, consider the necessity for the substance to be tested
for skin sensitization potential.
Review the Addendum for errors and omissions.
Provide conclusions and recommendations on the current
validation status of the LLNA applicability domain.
Does the information contained in the draft Addendum
support ICCVAM’s draft test method recommendations?
• 
• 
• 
The Panel concurred that that the data supported the
ICCVAM Test Method Recommendations for LLNA usefulness
and limitations.
The Panel considered all of the test materials as candidates
for testing in the LLNA, subject to the limitations outlined in
the ICCVAM Test Method Recommendations.
The Panel concluded that updated information did not
suggest the need for changes to recommendations for the
development of a revised standard method.
At the discretion of the testers, the Panel recommended the
inclusion of a suitable (representative) positive control from
the same category of materials to be tested (e.g., for testing
pesticides, select one representative positive control
pesticide).
The Panel concurred with ICCVAM’s recommendations for future studies, and concurred
that, before additional animal testing is conducted, consideration should be given to the
necessity for the substance to be tested for skin sensitization potential.
The complete LLNA Peer Review Panel Reports can be accessed at:
−  http://iccvam.niehs.nih.gov/docs/immunotox_docs/LLNAPRPRept2008.pdf
−  http://iccvam.niehs.nih.gov/docs/immunotox_docs/LLNAPRPRept2009.pdf
Michael Luster, Ph.D. (Panel Chair), Senior Consultant to the National Institute for
Occupational Safety and Health, Morgantown, WV
Nathalie Alépée, Ph.D., L’Oreal Research and Development, Aulnay sous Bois, France
Anne Marie Api, Ph.D., Research Institute for Fragrance Materials, Woodcliff Lake, NJ
Nancy Flournoy, M.S., Ph.D., University of Missouri – Columbia, Columbia, MO
Thomas Gebel, Ph.D., Federal Institute for Occupational Safety & Health, Dortmund,
Germany
Sidney Green, Ph.D., Howard University, Washington, DC
Kim Headrick, B.Admin., B.Sc., Health Canada, Ottawa, Ontario, Canada
Dagmar Jírová, M.D., Ph.D., National Institute of Public Health, Prague, Czech Republic
David Lovell, Ph.D., University of Surrey, Guilford, United Kingdom
Howard Maibach, M.D., University of California – San Francisco, San Francisco, CA
James McDougal, Ph.D., Wright State University, Dayton, OH
Michael Olson, Ph.D., GlaxoSmithKline, Research Triangle Park, NC
Raymond Pieters, Ph.D., Utrecht University, Utrecht, The Netherlands
Jean Regal, Ph.D., University of Minnesota Medical School, Duluth, MN
Jon Richmond, M.D., Home Office, London, United Kingdom
Peter Theran, V.M.D., Consultant, Massachusetts Society for the Prevention of Cruelty to
Animals, Novato, California
Stephen Ullrich, Ph.D., M.D. Anderson Cancer Center, Houston, TX
Michael Woolhiser, Ph.D., Dow Chemical, Midland, MI
Takahiko Yoshida, M.D., Ph.D., Asahikawa Medical College, Hokkaido, Japan
Boverhof D, Wiescinski C, Botham P, Lees D, Debruyne E, Repetto-Larsay M, et al. 2008. Interlaboratory
validation of 1% Pluronic® L92 surfactant as a suitable, aqueous vehicle for testing pesticide
formulations using the murine local lymph node assay. Toxicol Sci 105:79-85.
Agency for Toxic Substances and
Disease Registry
★ Moiz Mumtaz, Ph.D.
Bruce Fowler, Ph.D.
Ed Murray, Ph.D.
Eric Sampson, Ph.D.
Consumer Product Safety Commission
★ Marilyn L. Wind, Ph.D. (Chair)
◊ Kristina Hatlelid, Ph.D.
Joanna Matheson, Ph.D.
Adrienne Layton, Ph.D.
Department of Agriculture
★ Jodie Kulpa-Eddy, D.V.M. (Vice-chair)
◊ Elizabeth Goldentyer, D.V.M.
Department of Defense
★ Robert E. Foster, Ph.D.
◊ Patty Decot
Peter Schultheiss, D.V.M., DACLAM
Harry Salem, Ph.D.
Department of Energy
★ Michael Kuperberg, Ph.D.
◊ Marvin Stodolsky, Ph.D.
Department of the Interior
★ Barnett A. Rattner, Ph.D.
Department of Transportation
★George Cushmac, Ph.D.
◊ Steve Hwang, Ph.D.
1
The CPSC nomination may be viewed on the NICEATM-ICCVAM website at http://iccvam.niehs.nih.gov/methods/
immunotox/llnadocs/CPSC_LLNA_nom.pdf.
2
The report of the 2008 Peer Review Panel meeting is available at: http://iccvam.niehs.nih.gov/docs/
immunotox_docs/LLNAPRPRept2008.pdf
Health Effects Division
★ Jack Fowle, Ph.D.
3
The report of the 2009 Peer Review Panel meeting is available at: http://iccvam.niehs.nih.gov/docs/
immunotox_docs/LLNAPRPRept2009.pdf
Office of Pesticide Programs
◊Vicki Dellarco, Ph.D.
◊Tina Levine, Ph.D.
Deborah McCall
OECD Test Guidelines Program
Christine Augustyniak, Ph.D.
Final transmittal of these recommendations to agencies is currently in process.
•
• 
• 
ICCVAM recommends that the updated LLNA test method protocol (Appendix A, ICCVAM
2009a) should be used for all future LLNA studies, as it reduces animal use by 20%
compared to the 1999 ICCVAM-recommended protocol. Figure 1 shows a schematic of the
ICCVAM-recommended LLNA test method protocol.
If no dose-response information is required or there is no basis to believe that the test article
may be a sensitizer, a reduced LLNA test method protocol (testing only the high dose)
should be considered, which will further reduce animal use by up to 40% (ICCVAM 2009b).
• 
• 
• 
• 
1/1
•
Charge to the Peer Review Panel
• 
As indicated in Table 1, for many substances, there is a greater likelihood of
obtaining a positive result in the LLNA than in a GP test. Therefore, the potential for
possible overclassification may be a limitation of the LLNA.
Federal agencies should assess how well the test materials and findings in the
Addendum represent their substances of interest, particularly with respect to
chemical classes and potential biological effects.
Date
Abbreviations: GP = guinea pig skin sensitization outcomes; LLNA = murine local lymph node assay; No. = number.
Accuracy (concordance) = the proportion of correct outcomes (positive and negative) of a test method; Sensitivity = the
proportion of all positive substances based on results from the reference test method (i.e., guinea pig or human testing/
experience) that are classified as positive in the test method under evaluation (i.e. LLNA);
False negative rate = the proportion of all positive substances based on results in the reference test method (i.e., guinea pig
or human testing/experience) that are identified as negative in the test method under evaluation (i.e. LLNA);
Specificity = the proportion of all negative substances based on results from the reference test method (i.e., guinea pig or
human testing/experience) that are classified as negative in the test method under evaluation (i.e. LLNA);
False positive rate = the proportion of all negative substances based on results from the reference test method (i.e., guinea
pig or human testing/experience) that are identified as positive in the test method under evaluation (i.e. LLNA)
1 n = Number of substances included in this analysis.
2 The data on which the percentage calculation is based.
3 GP refers to outcomes obtained by studies conducted using either the guinea pig maximization test or the Buehler test.
4 Human refers to outcomes obtained by studies conducted using the human maximization test or the inclusion of the test
substance in a human patch test allergen kit.
•
Consumer Product Safety Commission
Joanna Matheson, Ph.D. (IWG Co-chair)
Marilyn Wind, Ph.D.
6
Final transmittal of these recommendations to agencies is
currently in process.
Public meetings of an international independent scientific peer
review panel (“Panel”) organized by ICCVAM and NICEATM
were held at U.S. Consumer Product Safety Commission
Headquarters in Bethesda, MD, on March 4-6, 2008, and at
the National Institutes of Health in Bethesda, MD, on April
28-29, 2009.
Peer Review Panel Conclusions
Dyes
LLNA vs. GP3
In conjunction with ECVAM and JaCVAM, ICCVAM has
developed internationally-harmonized test method performance
standards for the LLNA (ICCVAM 2009a) to evaluate the
performance of LLNA test methods that incorporate specific
protocol modifications (e.g., procedures to measure lymphocyte
proliferation) compared to the traditional LLNA.
Environmental Protection Agency
Food and Drug Administration
Office of the Commissioner
★Suzanne Fitzpatrick, Ph.D., DABT
Center for Drug Evaluation and Research
◊ Abigail C. Jacobs, Ph.D.
Paul C. Brown, Ph.D.
Center for Devices and Radiological Health
Melvin E. Stratmeyer, Ph.D.
Vasant Malshet, Ph.D., DABT
Center for Biologics Evaluation and
Research
Richard McFarland, Ph.D., M.D.
Ying Huang, Ph.D.
Center for Food Safety and Nutrition
David G. Hattan, Ph.D.
Robert L. Bronaugh, Ph.D.
Center for Veterinary Medicine
Devaraya Jagannath, Ph.D.
M. Cecilia Aguila, D.V.M.
National Center for Toxicological Research
Paul Howard, Ph.D.
Donna Mendrick, Ph.D.
Office of Regulatory Affairs
Lawrence A. D’Hoostelaere, Ph.D.
National Cancer Institute
Haneke KE, Tice RR, Carson BL, Margolin BH, Stokes WS. 2001. ICCVAM evaluation of the murine local
lymph node assay: III. Data analyses completed by the national toxicology program interagency
center for the evaluation of alternative toxicological methods. Regulatory Toxicology and
Pharmacology 34(3): 274-286.
ICCVAM 1999. The murine local lymph node assay: A test method for assessing the allergic contact
dermatitis potential of chemical/compounds. NIH Publication No. 99-4494. Research Triangle
Park, NC: National Institute of Environmental Health Sciences.
ICCVAM. 2009a. Recommended Performance Standards: Murine Local Lymph Node Assay. NIH
Publication Number 09-7357. Research Triangle Park, NC: National Institute of Environmental
Health Sciences. Available at http://iccvam.niehs.nih.gov/methods/immunotox/llna_PerfStds.htm.
ICCVAM. 2009b. The Reduced Murine Local Lymph Node Assay: An Alternative Test Method Using Fewer
Animals to Assess the Allergic Contact Dermatitis Potential of Chemicals and Products. NIH
Publication No. 09-6439. RTP, NC: NIEHS. Available: http://iccvam.niehs.nih.gov/methods/
immunotox/LLNA-LD/TMER.htm.
Sailstad DM, Hattan D, Hill RN, Stokes WS. 2001. ICCVAM evaluation of the murine local lymph node
assay: I. The ICCVAM review process. Regulatory Toxicology and Pharmacology 34(3): 249-257.
★T. Kevin Howcroft, Ph.D.
◊ Chand Khanna, D.V.M., Ph.D.
National Institute of Environmental
Health Sciences
★William S. Stokes, D.V.M., DACLAM
◊ Raymond R. Tice, Ph.D.
Rajendra S. Chhabra, Ph.D., DABT
Jerrold J. Heindel, Ph.D.
National Institute for Occupational
Safety and Health
★Paul Nicolaysen, V.M.D.
◊ K. Murali Rao, M.D., Ph.D.
National Institutes of Health
★Margaret D. Snyder, Ph.D.
★ Principal Agency Representative
◊ Alternate Principal Agency Representative
Dean JH, Twerdok LE, Tice RR, Sailstad DM, Hattan DG, Stokes WS. 2001. ICCVAM evaluation of the
murine local lymph node assay: II. Conclusions and recommendations of an independent
scientific peer review panel. Regulatory Toxicology and Pharmacology 34(3): 258-273.
National Library of Medicine
★Pertti (Bert) Hakkinen, Ph.D.
◊ Jeanne Goshorn, M.S.
Occupational Safety and Health
Administration
★Surender Ahir, Ph.D.
This poster was supported by the Intramural Research Program of the NIH, National Institute of
Environmental Health Sciences. ILS staff supported by NIEHS contract N01-ES 35504. The views
expressed above do not necessarily represent the official positions of any U.S. Federal agency.
Since the poster was written as part of the official duties of the authors, it can be freely copied.
ICCVAM and NICEATM gratefully acknowledge the following individuals and institutions that
submitted data to NICEATM used for the evaluation of the use of the LLNA to test pesticide
formulations and other products, metals, and substances in aqueous solutions.
Anne Marie Api, Ph.D.
Research Institute for Fragrance Materials
Woodcliff Lake, NJ
Michael Olson, Ph.D.
GlaxoSmithKline
Research Triangle Park, NC
Phil Botham, Ph.D.
European Crop Protection Association
Brussels, Belgium
Kirill Skirda, Ph.D.
TNO Quality of Life
Delft, Netherlands
Eric Debruyne, Ph.D.
Bayer CropScience SA, Sophia Antipolis
Cedex, France
Peter Ungeheuer, Ph.D.
European Federation for Cosmetic Ingredients
Frankfurt, Germany
G. Frank Gerberick, Ph.D.
Procter and Gamble Company
Cincinnati, OH
Michael Woolhiser, Ph.D.
Dow AgroSciences
Midland, MI
Dori Germolec, Ph.D.
National Toxicology Program
Research Triangle Park, NC